Reconstitution of T cell receptor repertoire after human allogeneic bone marrow transplantation

研究了人异基因骨髓移植后T细胞受体（TCR）库的重建。在从患者及其骨髓供体抽取血液样本之前获得知情同意书。通过使用接头连接PCR和微板杂交测定技术分析TCR V β和Valpha库。在供受者之间TCR库没有显著差异。移植8 -10周后，在所有检查的患者中观察到携带某些V β和Valpha亚家族的T细胞显著增加。移植1年后，扭曲的TCR库似乎恢复到正常模式。移植后CD 8 + CD 28-HLA-DR+T淋巴细胞数量增加，与TCR库变化相似。在该淋巴细胞亚群中优先使用频率增加的Vbeta和Valpha链。为了解骨髓移植后受者外周血淋巴细胞TCR的多样性，我们还对部分受者进行了CDR 3大小谱分析。在巨细胞病毒肺炎患者中观察到缺乏多个Vbeta亚家族和CDR 3复杂性的恢复受到干扰，这表明该方法可能有助于监测骨髓移植后的免疫重建。

Reconstitution of T cell receptor (TCR) repertoire after human allogeneic marrow transplantation was investigated. Informed consent was obtained before drawing blood samples from the patients and their marrow donors. TCR Vbeta and Valpha repertoire was analyzed by using adaptor ligation PCR and microplate hybridization assay techniques. There was no significant difference in TCR repertoire between donor-recipient. After8-10 weeks of transplant, a striking uncrease of the T cells carrying certain Vbeta and Valpha subfamilies was observed in all patients examined. Distorted TCR repertoire appears to retum to the normal pattem after 1 year of transplant. With the similar kinetics to the TCR repertoire changes, there was an increase of CD8+CD28-HLA-DR+T lymphocytes after transplant. Vbeta and Valpha chains with increased frequency were preferentially used in this lymphocyte subset. Taken together, the proliferation of CD8+CD28-HLA-DR+T lymphocytes results in the distortion of TCR repertoire after transplant.To investigate the TCR diversity of blood lymphocytes in marrow recipients, CDR3 size spectratyping analsis was also performed in several patients. The lack of multiple Vbeta subfamilies and the disturbed recovery of CDR3 complexity were observed in the patient with cytomegalovirus pneumonia, suggesting that this method might be useful for monitoring immune reconstitution after marrow transplantation.